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Related Experiment Videos

Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms.

K Gerritse1, P de Knijff, G van Ierssel

  • 1Medical Biological Laboratory TNO, Rijswijk, The Netherlands.

Journal of Lipid Research
|February 1, 1992
PubMed
Summary

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Researchers developed a specific monoclonal antibody targeting apolipoprotein E2 (apoE2) with a cysteine mutation. This antibody enables a novel immunoblot assay for detecting apoE2 in human sera without complex pre-processing.

Area of Science:

  • Biochemistry
  • Immunology
  • Medical Diagnostics

Background:

  • Apolipoprotein E (apoE) plays a crucial role in lipid metabolism and is associated with various diseases.
  • Specific isoforms of apoE, such as apoE2 with an arginine to cysteine substitution at position 158 (apoE2(Arg158----Cys)), have distinct functional properties.
  • Detecting specific apoE variants in human serum is important for clinical diagnostics but often requires laborious methods like isoelectric focusing.

Purpose of the Study:

  • To develop a specific monoclonal antibody (MAb) targeting the apoE2(Arg158----Cys) variant.
  • To establish a sensitive, rapid, and straightforward immunoblot assay for detecting apoE2(Arg158----Cys) in human serum.

Main Methods:

  • Immunization of mice with a synthetic peptide (amino acids 154-172) homologous to human and mouse apolipoprotein E.

Related Experiment Videos

  • Selection of a peptide with a surprisingly low predicted antigenicity index that elicited a specific immune response.
  • Generation and characterization of a variant-specific anti-peptide MAb recognizing apoE2(Arg158----Cys) but not apoE3.
  • Development of an immunoblot assay utilizing the generated MAb for direct detection of apoE2(Arg158----Cys) in serum.

    • Main Results:

    • A specific monoclonal antibody (MAb) was successfully raised against the apoE2(Arg158----Cys) variant using a specific synthetic peptide.
    • The generated MAb demonstrated high specificity, recognizing apoE2(Arg158----Cys) while showing no cross-reactivity with apoE3.
    • A novel immunoblot assay was developed, proving to be sensitive, time-saving, and simple for detecting apoE2(Arg158----Cys) in human sera without prior isoelectric focusing.

    Conclusions:

    • A specific monoclonal antibody against apoE2(Arg158----Cys) can be generated using a low-antigenicity peptide.
    • The developed MAb and subsequent immunoblot assay offer a significant improvement for the detection of apoE2(Arg158----Cys) in clinical settings.
    • This method simplifies the diagnostic process for apoE2(Arg158----Cys) related conditions.